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Direct interaction of post‐synaptic density‐95/Dlg/ZO‐1 domain‐containing synaptic molecule Shank3 with GluR1 α‐amino‐3‐hydroxy‐5‐methyl‐4‐isoxazole propionic acid receptor
Author(s) -
Uchino Shigeo,
Wada Hidenori,
Honda Shizuyo,
Nakamura Yasuko,
Ondo Yumiko,
Uchiyama Takayoshi,
Tsutsumi Moe,
Suzuki Eri,
Hirasawa Takae,
Kohsaka Shinichi
Publication year - 2006
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.2006.03831.x
Subject(s) - pdz domain , ampa receptor , protein subunit , postsynaptic density , biology , microbiology and biotechnology , scaffold protein , glutamate receptor , receptor , dendritic spine , synaptic plasticity , biochemistry , hippocampal formation , neuroscience , signal transduction , gene
A class of scaffolding protein containing the post‐synaptic density‐95/Dlg/ZO‐1 (PDZ) domain is thought to be involved in synaptic trafficking of α‐amino‐3‐hydroxy‐5‐methyl‐4‐isoxazole propionic acid (AMPA) receptors during development. To clarify the molecular mechanism of AMPA receptor trafficking, we performed a yeast two‐hybrid screening system using the cytoplasmic tail of the GluR1 subunit of AMPA receptor as a bait and identified a synaptic molecule, Shank3/ProSAP2, as a GluR1 subunit‐interacting molecule. Shank3 is a PDZ domain‐containing multidomain protein and is predominantly expressed in developing neurons. Using the glutathione S‐transferase pull‐down assay and immunoprecipitation technique we demonstrated that the GluR1 subunit directly binds to the PDZ domain of Shank3 via its carboxyl terminal PDZ‐binding motif. We raised anti‐Shank3 antibody to investigate the expression of Shank3 in cortical neurons. The pattern of Shank3 immunoreactivity was strikingly punctate, mainly observed in the spines, and closely matched the pattern of post‐synaptic density‐95 immunoreactivity, indicating that Shank3 is colocalized with post‐synaptic density‐95 in the same spines. When Shank3 and the GluR1 subunit were overexpressed in primary cortical neurons, they were also colocalized in the spines. Taken together with the biochemical interaction of Shank3 with the GluR1 subunit, these results suggest that Shank3 is an important molecule that interacts with GluR1 AMPA receptor at synaptic sites of developing neurons.